Cracking the Infection Control Code: Using and Interpreting Diagnostic Tests to Control Infectious Diseases in Shelters Dr. Ronald D. Schultz, Professor and Chair Department of Pathobiological Sciences School of Veterinary Medicine University of Wisconsin-Madison, WI 53706 Important Tools for Disease Outbreak Investigation and Control 1. Observations 2. Testing/Diagnostics 3. Prevention/Treatment Diagnostics: Two Basic Methods (Immunity vs. Infection) 1. Detect Ab Immunity (exceptions*) 2. Detect Ag or Nucleic Acid (Virus, Bacteria, Parasite, other) Infection 3. Method of detection * FIV, MAT - Lepto 1
Diagnostics 1. Methods to Detect Agent (Infection) A. 1. Immunologic Ag Capture (examples: FeLV, fecal parvovirus) 2. Immunologic Antibody (e.g. FIV) B. Molecular PCR* (examples: CDV, CPV-2, FHV-1, FPV, FCV, Leptospira, Bordetella) * Positive may not show infectious agent is present in samples, but does show the nucleic acid is present. Diagnostics 2. Methods to Detect Antibody (Infection or Immunity) A. Virus Neutralization (VN, SN): CDV, FCV, FHV-1, CAV-1,2 - Immunity B. Hemagglutination Inhibition (HI): CPV-2, FPV, CIV- Immunity C. Agglutination (eg MAT): Leptospira D. ELISA FIV, CPV/FPV, FeLV Infection E. Fluorescence Lyme immunity and infection Diagnostics PCR for Infectious Agents Many shelters and practices are having samples tested by this method. Excellent test, as it has high sensitivity and specificity! (Very New = Quality Control Lacking) Idexx Lab and various State Diagnostic Laboratories Interpretetation of Results? Cost? Very Expensive Test 2
Diagnostics: PCR How to interpret results: Sensitivity Cycle No. 35: pos > 35 39: suspect 40: neg (These values vary somewhat among laboratories that perform the PCR.) The lower the cycle number, the more positive the result! How Does Vaccination Affect the Results of PCR Testing? We and hopefully others are now and have been doing studies to try to answer this question! Ideally, collect samples prior to vaccination with infectious (modified live) vaccines! PCR Testing (cont.) Our results suggest: Samples collected after vaccination can be positive for up to 2 or more weeks*, depending on immune status of animal at time of vaccination and the vaccine. * Fecal samples for CPV-2 or FPV may be pos for more than 2 weeks! Use clinical signs of parvovirus infection and fecal SNAP test for diagnosis of CPV- 2 or FPV, not PCR in animals vaccinated during previous 3 weeks. 3
PCR Testing (cont.) CPV-2/FPV, CDV, CAV-2, FHV-1, FCV vaccinated* dogs and cats positive for 2 or more (CPV-2/FPV) weeks when MLV vaccines are used. *rcdv and noninfectious vaccine do not cause the animal to become PCR Pos.! PCR Testing (cont.) Be very careful when collecting samples, as you can readily cross contaminate samples. Contamination from another animal, from a sample, from a vaccine! Remember PCR is very sensitive! PCR (cont.) When Collecting samples for PCR, be very careful to not contaminate samples e.g. change gloves Use PCR to ensure animal is no longer infected and shedding 4
Infectious Diseases Commonly Seen in Shelters Canine and Feline Infectious Respiratory Disease Complexes ( Canine and Feline Cold ) PCR is of limited value as most animal will be Positive! Canine Diseases Canine Distemper (CDV) Vomiting/Diarrhea/Respiratory Canine Parvovirus (CPV-2) Vomiting/Diarrhea PCR is of value! Ag Fecal Parvo Snap Feline Diseases Feline Parvovirus (Feline Panleukopenia) Vomiting/Diarrhea PCR is of value! Ag Fecal Parvo Snap 5
Miscellaneous Diseases/Infections To Test For: Idexx Feline Combo Feline Leukemia Virus (FeLV) Ag Feline Immunodeficiency Virus Ab (FIV) Idexx 4DX Heartworm Ag Ehrlichia Ab Anaplasma Ab Lyme (Borrelic) Ab (C6) Idexx Fecal Parvo Snap What Do The Test Results Tell You? Dog That is CDV Positive Antibody (Ab) Immune PCR Infected Ab & PCR Virus was or is present and antibody has developed Virus may be neutralized and not infectious! How to Reduce Costs When Using PCR Pool Samples Nasal swabs CDV Fecal swabs CPV-2, FPV 6
Dogs Infected with CDV Dogs with signs of kennel cough may be infected with CDV! CDV can cause kennel cough like condition! Incubation period can be up to 6 weeks! PCR Positive CDV Incubation Period up to 6 weeks PCR Pos. no Clinical Signs Cats and Dogs with FPV or CPV Incubation period is usually 3 to 4 days. Can be up to 7 days! PCR - positive 7
FPV/CPV Incubation period 3 to 7 days PCR Pos. no Clinical Signs PCR Positive Animals CDV, CPV-2, FPV Do not adopt these animals Hold them until PCR Neg. Can Use PCR to ensure animal is no longer infected and shedding When collecting samples for PCR, be very careful to not cross contaminate samples e.g. change gloves Very sensitive test Antibody Testing Can Be Performed By a Diagnostic Lab or In-House Tests are Available Diagnostic labs often perform serologic tests that provide an antibody titer such as serum neutralization tests for CDV, CAV-1, rabies or hemagglutination inhibition (HI) tests for CPV-2, FPV. In-house tests are often ELISA based. 8
Virus Neutralization Test HI Test In House (On Site) Tests 1. Titer CHEK - CDV and CPV-2 ELISA 96 well plate, multiple tests 2. VacciCheck - CDV, CPV-2, CAV-1 Dog; FPV, FCV, FHV-1 Cat - ELISA, Immunocomb, individual or multiple tests 9
On Site Tests There are two commercial inoffice or on-site ELISA tests, TiterCHEK and VacciCheck that have been correlated with serum samples from dogs that resisted challenge with CDV and CPV-2 and also with the Gold Standard serologic tests On Site Tests TiterCHEK and VacciCheck Very useful in puppies to ensure that there was an immune response to CDV/CPV-2 after their series of vaccinations If serum sample is collected 2 or more weeks after the last puppy vaccination the test should be positive for both viruses TiterCHEK and VacciCheck When the test is not positive, revaccinate and test again If the test is not positive after revaccination, further testing using the Gold Standard Test may be required 10
TiterCHEK and VacciCheck A negative shows there is no antibody or it is lower than the minimum amount that can be detected Titerchek - http://www.zoetis.com/ VacciCheck - http://vaccicheck.com/ Antibody Testing Serum antibody correlates with protection from the following diseases: Dog: CAV-1, CDV, CPV-2, Rabies, Lyme (borreliacidal Ab) Cat: FPV, FCV?, Rabies Antibody Response to Vaccines The detection of antibody as a result of vaccination with an infectious vaccine (MLV, attenuated, viral vectored recombinant) demonstrates development of B and T cell immunity! All complex antigens require T cell help to get a B cell to differentiate and produce antibody. 11
Antibody Testing (cont) When using antibody tests to demonstrate vaccinal immunity, they are useful: 1. To demonstrate immunity after completing the puppy or kitten series. Take a blood sample 2 or more weeks after last vaccination at 14 to 16 weeks or older. If the test shows antibody is present, the animal has developed immunity! Antibody Testing (cont) 2. To demonstrate the animal is immune one year or more after vaccination I don t recommend retesting more often than every 3 years after demonstrating the animal is positive at one year. How Can Antibody Testing be Used in Kennels/Shelters? Shelters having significant problems with dogs getting infected with CDV or CPV-2 or cats getting infected with FPV, antibody testing and vaccination at admission are important in controlling and/or eliminating the problem! 12
Outbreak Control (Kennels/Shelters) For effective control of infectious disease, vaccinate with the core vaccines immediately upon entry or, when possible, prior to entry, for dogs and cats that are not antibody positive. Outbreak Control (Kennels/Shelters) Infectious disease outbreaks caused by CDV, CPV-2, or FPV can be prevented and they can be controlled without euthanizing large numbers of dogs or cats. What Does it Mean Ab Pos. + PCR Pos. Virus was (is) present and antibody is present Virus may be neutralized and will not infect nor cause disease 13
Outbreaks PCR Pos. Animals CDV, CPV-2, FPV Do not adopt these animals. Keep them until PCR Neg. How to Reduce Costs When Using PCR Pool Samples Nasal swabs CDV Fecal swabs CPV-2, FPV Vaccines that Every Shelter Animal Should Receive at or Prior to Admission Canine: (Shelter Core Vaccines) CDV/CPV-2/CAV-2/CPI-5 (Combo) Bb/CPI IN or Bb IN or Oral Feline: (Shelter Core Vaccines) FPV/FCV/FVR Canine & Feline: Receive at Adoption Rabies Vaccine 3 yr. product 14
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